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J Econ Entomol

Title:		A Real-Time PCR Assay for the Separation of Autographa gamma (Noctuidae: Plusiinae) From Morphologically Similar Species in North America
Author(s):		Tembrock LR; Farris RE; Ledezma L; Barr NB; Gilligan TM;
Address:		"Department of Bioagricultural Sciences and Pest Management, Colorado State University. USDA-APHIS-PPQ-Science & Technology, Mission Laboratory. USDA-APHIS-PPQ-Science & Technology, Identification Technology Program"
Journal Title:		J Econ Entomol
Year:		2017
Volume:		110
Issue:		6
Page Number:		2609 - 2617
DOI:		10.1093/jee/tox256
ISSN/ISBN:		1938-291X (Electronic) 0022-0493 (Linking)
Abstract:		"The silver Y moth, Autographa gamma L. (Noctuidae: Plusiinae), is a pest of major economic importance in its native range of Europe, Asia, and North Africa. Although not present in North America, larvae of A. gamma are commonly intercepted in commodity shipments at U.S. ports, and adult surveys are conducted each year in more than 20 states. Because of the similarity of A. gamma to several native North American species that are attracted to the same pheromone lure, morphological identification of adults is difficult and requires dissection. In 2010, a specimen of Autographa californica (Speyer, 1875) (Lepidoptera: Noctuidae) from Pennsylvania was incorrectly identified as A. gamma, signaling the need for an alternative method of rapid identification. Here we detail a real-time PCR assay capable of identifying A. gamma specimens in approximately 45 min using extracted DNA. The assay uses a hydrolysis probe that targets a species-specific segment of the CO1 DNA barcode region, while a control probe targets a conserved region of 18S rDNA. The assay was tested with two independent runs of 452 specimens of Plusiinae representing 23 different species. The assay provided unambiguous data 99.7% of the time and did not result in any false positives; these data were used to develop a rule set for interpreting the real-time PCR results. In addition, the same diagnostic probe was tested in bulk sample simulations using real-time PCR and droplet digital PCR where A. gamma could be detected in concentrations as low as 1:1,000,000 (gamma:californica). These experiments provide baseline data for developing a bulk sample assay"
Keywords:		"Animals DNA Barcoding, Taxonomic Electron Transport Complex IV/genetics Insect Proteins/genetics *Introduced Species Larva/genetics/growth & development Moths/*genetics/growth & development North America Real-Time Polymerase Chain Reaction/*methods Sensit;"
Notes:		"MedlineTembrock, Luke R Farris, Roxanne E Ledezma, Lisa Barr, Norman B Gilligan, Todd M eng England 2017/10/14 J Econ Entomol. 2017 Dec 5; 110(6):2609-2617. doi: 10.1093/jee/tox256"